Exploring the treatment of sepsis-associated acute lung injury with Liangge Powder via ERK1/2 and PI3K/AKT pathways: based on network pharmacology and whole animal experimentation / 中华劳动卫生职业病杂志

Objective: To investigate the therapeutic effect and mechanism of Liangge Powder against sepsis-induced acute lung injury (ALI) . Methods: From April to December 2021, the key components of Liangge Powder and its targets against sepsis-induced ALI were analyzed by network pharmacology, and to enrich for relevant signaling pathways. A total of 90 male Sprague-Dawley rats were randomly assigned to sham-operated group, sepsis-induced ALI model group (model group), Liangge Powder low, medium and high dose group, ten rats in the sham-operated group and 20 rats in each of the remaining four groups. Sepsis-induced ALI model was established by cecal ligation and puncture. Sham-operated group: gavage with 2 ml saline and no surgical treatment. Model group: surgery was performed and 2 ml saline was gavaged. Liangge Powder low, medium and high dose groups: surgery and gavage of Liangge Powder 3.9, 7.8 and 15.6 g/kg, respectively. To measure the wet/dry mass ratio of rats lung tissue and evaluate the permeability of alveolar capillary barrier. Lung tissue were stained with hematoxylin and eosin for histomorphological analysis. The levels of tumor necrosis factor-alpha (TNF-α), interleukin (IL) -6 and IL-1β in bronchoalveolar lavage fluid (BALF) were measured by enzyme-linked immunosorbent assay. The relative protein expression levels of p-phosphatidylinositol 3-kinase (PI3K), p-protein kinase B (AKT), and p-ertracellular regulated protein kinases (ERK) were detected via Western blot analysis. Results: Network pharmacology analysis indicated that 177 active compounds of Liangge Powder were selected. A total of 88 potential targets of Liangge Powder on sepsis-induced ALI were identified. 354 GO terms of Liangge Powder on sepsis-induced ALI and 108 pathways were identified using GO and KEGG analysis. PI3K/AKT signaling pathway was recognized to play an important role for Liangge Powder against sepsis-induced ALI. Compared with the sham-operated group, the lung tissue wet/dry weight ratio of rats in the model group (6.35±0.95) was increased (P<0.001). HE staining showed the destruction of normal structure of lung tissue. The levels of IL-6 [ (392.36±66.83) pg/ml], IL-1β [ (137.11±26.83) pg/ml] and TNF-α [ (238.34±59.36) pg/ml] were increased in the BALF (P<0.001, =0.001, <0.001), and the expression levels of p-PI3K, p-AKT and p-ERK1/2 proteins (1.04±0.15, 0.51±0.04, 2.31±0.41) were increased in lung tissue (P=0.002, 0.003, 0.005). The lung histopathological changes were reduced in each dose group of Liangge Powder compared with the model group. Compared with the model group, the wet/dry weight ratio of lung tissue (4.29±1.26) was reduced in the Liangge Powder medium dose group (P=0.019). TNF-α level [ (147.85±39.05) pg/ml] was reduced (P=0.022), and the relative protein expression levels of p-PI3K (0.37±0.18) and p-ERK1/2 (1.36±0.07) were reduced (P=0.008, 0.017). The wet/dry weight ratio of lung tissue (4.16±0.66) was reduced in the high-dose group (P=0.003). Levels of IL-6, IL-1β and TNF-α[ (187.98±53.28) pg/ml, (92.45±25.39) pg/ml, (129.77±55.94) pg/ml] were reduced (P=0.001, 0.027, 0.018), and relative protein expression levels of p-PI3K, p-AKT and p-ERK1/2 (0.65±0.05, 0.31±0.08, 1.30±0.12) were reduced (P=0.013, 0.018, 0.015) . Conclusion: Liangge Powder has therapeutic effects in rats with sepsis-induced ALI, and the mechanism may be related to the inhibition of ERK1/2 and PI3K/AKT pathway activation in lung tissue.

T lymphocyte subsets combined with inflammatory indicators for auxiliary diagnosis and prognosis evaluation of sepsis / 中华预防医学杂志

Objective: To explore the application value of T lymphocyte subsets combined with procalcitonin (PCT), C-reactive protein (CRP), neutrophil to lymphocyte ratio (NLR) and white blood cell count (WBC) in the auxiliary diagnosis and prognosis evaluation of sepsis. Methods: In a retrospective study, seventy-two patients with sepsis diagnosed and treated in Tianjin First Central Hospital from June 2018 to April 2021 were selected as the research objects, and included in the sepsis group were 46 males and 26 females, aged 68 (57.3, 80.3) years. In addition, 111 patients with local infection admitted to hospital during the same period were included in the local infection group, including 62 males and 49 females, aged 68 (51, 77) years. Sepsis patients were divided into survival group (43 cases) and death group (29 cases) according to the 28-day outcome. CD3+, CD4+, CD8+, CD4+/CD8+ ratio were detected by flow cytometry within 24 h after admission, PCT was detected by ELISA, CRP was detected by immunoturbidimetry, blood routine examination, blood lactic acid (Lac) and oxygen partial pressure (PO2) were detected by instrumental method. Multivariate Logistic regression analysis was used to evaluate the correlation between each indicator and sepsis, and receiver operating characteristic curve (ROC) was drawn to evaluate the diagnostic value of each indicator for sepsis. Multivariate Logistic regression analysis and Kaplan Meier survival analysis were used to evaluate the prognostic value of each index for patients with sepsis. Results: Peripheral blood CD3+, CD4+, CD8+, CD4+/CD8+ ratio and PLT in sepsis group were significantly lower than those in local infection group(Z=-8.184,P<0.001;Z=-7.210,P<0.001;Z=-5.936,P<0.001;Z=-2.700,P=0.007;Z=-6.381,P<0.001); PCT, CRP, NLR and Lac levels were significantly higher than those in local infection group(Z=-8.262,P<0.001;Z=-3.094,P=0.002;Z=-9.004,P<0.001;Z=-4.770,P<0.001). Multivariate Logistic regression model showed that PCT, NLR, CD3+, CD8+, CD4+/CD8+ were independent risk factors for sepsis. According to ROC curve analysis, AUC of sepsis patients diagnosed by each indicator were 0.862, 0.894, 0.858, 0.760 and 0.618, respectively. The cut-off values were 3.075 ng/ml, 10.715, 44.935×109/L, 27.463×109/L and 0.750, respectively. The NLR sensitivity was 80.6%, and the CD3+ specificity was 94.6%. The AUC of combined detection of PCT and NLR was 0.947, sensitivity was 87.5% and specificity was 91.9%. The combined detection AUC of PCT, NLR, CD3+, CD4+/CD8+ was 0.958, the sensitivity and specificity were 90.3% and 91.0% respectively(P<0.001). PCT and Lac in death group were significantly higher than those in survival group(Z=-2.302,P=0.021;Z=-3.095,P=0.002);Peripheral blood CD4+/CD8+ levels were significantly lower than those in survival group(Z=-3.691,P<0.001),Multivariate Logistic regression model showed that CD4+/CD8+ ratio was an independent risk factor for 28 d mortality in patients with sepsis (P<0.001). The ROC curve showed that the AUC was 0.758, and the Youden index reached the maximum when the cut-off value was 1.27, the sensitivity and specificity were 79.3% and 60.5%, respectively. Compared with patients with CD4+/CD8+ ≥1.27, 28-day mortality was significantly increased in patients with CD4+/CD8+<1.27 (P=0.032). Conclusion: The combined detection of PCT, NLR, CD3+ and CD4+/CD8+ can improve the auxiliary diagnostic efficiency of sepsis, and the ratio of CD4+/CD8+ in peripheral blood may have certain predictive value for the prognosis of sepsis.

Inhibitory effects of petasin on human colon carcinoma cells mediated by inactivation of Akt/mTOR pathway / 中华医学杂志(英文版)

BACKGROUND@#Colorectal cancer is the third most common cancer worldwide and still lack of effective therapy so far. Petasin, a natural product found in plants of the genus Petasites, has been reported to possess anticancer activity. The present study aimed to investigate the anticolon cancer activity of petasin both in vitro and in vivo. The molecular mechanism of petasin was also further explored.@*METHODS@#Caco-2, LoVo, SW-620, and HT-29 cell lines were used to detect the inhibitory effect of petasin on colon cancer proliferation. Cell viability was determined using the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay. Cell apoptosis was analyzed by flow cytometry. Hoechst 33258 staining was used to visualize morphological changes. Cell migration was assessed using a wound-healing migration assay, and cell invasion was investigated using Transwell chambers. Western blotting assays were employed to evaluate the expression levels of proteins in the protein kinase B/mammalian target of rapamycin (Akt/mTOR) signaling pathway. Finally, in vivo activity of petasin was evaluated using the SW-620 subcutaneous tumor model established in Balb/c nude mice. Twelve rats were randomly divided into control group and 10 mg/kg petasin group. The tumor volume was calculated every 7 days for 28 days. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay was performed to assess the apoptotic effect of petasin. Differences between two groups were assessed by analysis of independent-sample t tests.@*RESULTS@#Petasin significantly inhibited the proliferation of human colon carcinoma cell lines, induced apoptosis, and suppressed migration and invasion in SW-620 cells. Western blotting results showed that petasin decreased the phosphorylation of Akt (1.01 ± 0.16 vs. 0.74 ± 0.06, P = 0.042), mTOR (0.71 ± 0.12 vs. 0.32 ± 0.11, P = 0.013), and P70S6K (1.23 ± 0.21 vs. 0.85 ± 0.14, P = 0.008), elevated the expression of caspase-3 (0.41 ± 0.09 vs. 0.74 ± 0.12, P = 0.018) and caspase-9 (1.10 ± 0.27 vs. 1.98 ± 0.22, P = 0.009), decreased the Bcl-2 protein (2.75 ± 0.47 vs. 1.51 ± 0.36, P = 0.008), downregulated the expression of matrix metalloproteinase (MMP)-3 (1.51 ± 0.31 vs. 0.82 ± 0.11, P = 0.021) and MMP-9 (1.56 ± 0.32 vs. 0.94 ± 0.15, P = 0.039) in SW-620 cell. In vivo, 10 mg/kg petasin inhibited tumor growth in Balb/c nude mice (924.18 ± 101.23 vs. 577.67 ± 75.12 mm at day 28, P = 0.001) and induced apoptosis (3.6 ± 0.7% vs. 36.0 ± 4.9%, P = 0.001) in tumor tissues.@*CONCLUSIONS@#Petasin inhibits the proliferation of colon cancer SW-620 cells via inactivating the Akt/mTOR pathway. Our findings suggest petasin as a potential candidate for colon cancer therapy.

Inhibitory effects of petasin on human colon carcinoma cells mediated by inactivation of Akt/mTOR pathway / 中华医学杂志(英文版)

Background:@#Colorectal cancer is the third most common cancer worldwide and still lack of effective therapy so far. Petasin, a natural product found in plants of the genus Petasites, has been reported to possess anticancer activity. The present study aimed to investigate the anticolon cancer activity of petasin both in vitro and in vivo. The molecular mechanism of petasin was also further explored.@*Methods:@#Caco-2, LoVo, SW-620, and HT-29 cell lines were used to detect the inhibitory effect of petasin on colon cancer proliferation. Cell viability was determined using the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay. Cell apoptosis was analyzed by flow cytometry. Hoechst 33258 staining was used to visualize morphological changes. Cell migration was assessed using a wound-healing migration assay, and cell invasion was investigated using Transwell chambers. Western blotting assays were employed to evaluate the expression levels of proteins in the protein kinase B/mammalian target of rapamycin (Akt/mTOR) signaling pathway. Finally, in vivo activity of petasin was evaluated using the SW-620 subcutaneous tumor model established in Balb/c nude mice. Twelve rats were randomly divided into control group and 10 mg/kg petasin group. The tumor volume was calculated every 7 days for 28 days. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay was performed to assess the apoptotic effect of petasin. Differences between two groups were assessed by analysis of independent-sample t tests.@*Results:@#Petasin significantly inhibited the proliferation of human colon carcinoma cell lines, induced apoptosis, and suppressed migration and invasion in SW-620 cells. Western blotting results showed that petasin decreased the phosphorylation of Akt (1.01 ± 0.16 vs. 0.74 ± 0.06, P = 0.042), mTOR (0.71 ± 0.12 vs. 0.32 ± 0.11, P = 0.013), and P70S6K (1.23 ± 0.21 vs. 0.85 ± 0.14, P = 0.008), elevated the expression of caspase-3 (0.41 ± 0.09 vs. 0.74 ± 0.12, P = 0.018) and caspase-9 (1.10 ± 0.27 vs. 1.98 ± 0.22, P = 0.009), decreased the Bcl-2 protein (2.75 ± 0.47 vs. 1.51 ± 0.36, P = 0.008), downregulated the expression of matrix metalloproteinase (MMP)-3 (1.51 ± 0.31 vs. 0.82 ± 0.11, P = 0.021) and MMP-9 (1.56 ± 0.32 vs. 0.94 ± 0.15, P = 0.039) in SW-620 cell. In vivo, 10 mg/kg petasin inhibited tumor growth in Balb/c nude mice (924.18 ± 101.23 vs. 577.67 ± 75.12 mm3 at day 28, P = 0.001) and induced apoptosis (3.6 ± 0.7% vs. 36.0 ± 4.9%, P = 0.001) in tumor tissues.@*Conclusions:@#Petasin inhibits the proliferation of colon cancer SW-620 cells via inactivating the Akt/mTOR pathway. Our findings suggest petasin as a potential candidate for colon cancer therapy.

Related research on corneal higher-order aberrations after different ways refractive surgery / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To evaluate the changes of corneal high - order aberration (including Coma, Spab, RMSh) after laser in situ keratomileusis (LASIK) with femtosecond laser, sub- Bowman keratomileusis ( SBK ) and laser epithelial keratomileusis (LASEK). METHODS: Of 82 myopic patients ( 164 eyes ), 31 patients (62 eyes) were treated by FS-LASIK, 31 patients (62 eyes) were treated by SBK, 20 patients (40 eyes) were treated by LASEK. Sirius system was used for measuring the coma aberration, spherical aberration, and high order aberration at 1, 15d,1, 3mo after surgery. RESULTS: 1) Vision: The uncorrected visual acuity of the three groups had no differences (P>0. 05). 2) Corneal aberrations: Three kinds of surgical procedure for patients with corneal aberration had significant impact. The C7, C8, C12 and RMSh of three groups were increased significantly (P<0. 05). The C7, C8, C12 and RMSh were not recovered to preoperative levels after 3mo. But the increase of patients after FS- LASIK was smaller than the other two groups, with statistical significance (P<0. 05). CONCLUSION: Compared with SBK and LASEK, FS - LASIK has better visual acuity in the early postoperative and corneal higher-order aberrations increase is relatively small.

Evaluation and clinical application of sacral s1 vestibule measurements in chinese adults / Evaluación y aplicación clínica de las mediciones del vestíbulo sacral s1 en adultos chinos

This study aims to measure the anatomic parameters of the sacral 1 (S1) vestibule in Chinese adults and to discuss their clinical application during iliosacral screw fixation for pelvic posterior ring injury. Three-dimensional computed tomography (CT) reconstructions were performed on 36 individuals, and the parameters of their S1 vestibules were measured. Vestibular width (VW) was 25.15±2.91 mm, vestibular height (VH) was 20.94±3.03 mm, and mean vestibular size (VS) was 400.23±85.11 mm2. The mean angle of superior inclination was 30.85°±9.22°, and the mean anterior inclination (AI) was 13.91°±6.25°. VW and VS were significantly smaller in females than in males (p<0.05), but no statistical differences were found between the left and right sides. The S1 vestibules of Chinese patients are smaller than those reported for Caucasians. Therefore, the placement of iliosacral screws should be considered carefully based on the size, gender, and ethnicity of the patient. The anatomic parameters of females were much smaller than those of males and close to the minimum requirement for fracture fixation. Therefore, female Chinese patients who need iliosacral screws should undergo preoperative CT scans to measure S1 vestibule parameters to make individual operational plans.

El estudio tuvo como objetivo medir los parámetros anatómicos del vestíbulo sacral 1 (S1) en individuos adultos chinos y discutir su aplicación clínica durante la fijación de tornillo iliosacral por lesiones del anillo pélvico posterior. Se realizaron reconstrucciones de tomografía computarizada tridimensional (TC) en 36 individuos y se midieron los parámetros de sus vestíbulos S1. El ancho vestibular (AV) fue 25,15±2,91 mm, la altura vestibular fue 20,94±3,03 mm y el tamaño medio vestibular (TV) fue 400,23±85,11 mm2. El ángulo medio de inclinación superior fue 30,85°±9,22° e inclinación anterior media fue 13,91°±6,25°. AV y TV fueron significativamente menores en las mujeres que en los hombres (p<0,05), sin embargo no se encontraron diferencias estadísticas entre los lados izquierdo y derecho. Los vestíbulos S1 de pacientes chinos son más pequeños que los reportados para los caucásicos. Por lo tanto, la colocación de tornillos iliosacros debe ser considerada cuidadosamente basada en el tamaño, el sexo y origen étnico del paciente. Los parámetros anatómicos de las mujeres eran significativamente más pequeños que los de los hombres y cercanos al requerimiento mínimo para la fijación de fracturas. Por lo tanto, pacientes de sexo femenino chino que requieren tornillos iliosacros deben ser sometidos a tomografías computarizadas preoperatorias para medir los parámetros del vestíbulo S1 a con el objetivo de programar un plan operativo individual.

Central corneal thickness using EX500 excimer laser workstation / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To evaluate the accuracy of central conreal thickness ( CCT ) using EX500 Excimer Laser workstation (EX500) in laser in situ keratomileusis (LASIK) patients.METHODS:The CCT of 120 eyes (63 patients) who had LASIK between January 2013 and June 2013 were measured by A- scan and EX500. Three groups were classified: >550μm, 500 ~550μm, RESULTS: The average preoperative CCT value was 527. 9±34. 3μm measured by A-scan, 528. 5±34. 6μm measured by EX500. There was no significant difference between these two measurements (t=1. 736, P=0. 085). In group which CCT >550μm, the average preoperative CCT value was 571. 4±17. 3μm measured by A-scan, 572.7±15. 7μm measured by EX500. There was no significant difference between these two measurements (t=1. 857, P=0. 072). In group which CCT 500 ~ 550μm, the average preoperative CCT value was 523. 4±13. 1μm measured by A-scan, 524. 2±12. 4μm measured by EX500. There was no significant difference between these two measurements ( t=1. 934, P = 0. 058 ). In group which CCT CONCLUSION: There is no significant difference between preoperative CCT value measured by A-scan and EX500. After corneal flap lifting and keratomileusis, the CCT value measured by EX500 is smaller than measured by A-scan.

Comparison of two surgical treatments for pediatric lower eyelid trichiasis / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To compare the clinical results of suture method and partial eyelash resection treating for pediatric eyelid trichiasis, and screen an effective method for the treatment of pediatric lower eyelid trichiasis. METHODS: Fifty-six cases of pediatric patients with lower eyelid trichiasis were randomly divided into a control group and an observation group in accordance with the method of drawing lots, and each group was 28 cases. The control group was treated with suture method, and the observation group was treated with partial eyelash resection. The clinical efficacy, patient satisfaction before and after treatment, and the incidence of complications were compared. RESULTS:(1) The clinically total effective rate was 74%of the control group, which was 89% of the observation group, and there were statistical differences of the clinical efficacy between the two groups ( P CONCLUSION: The treatment of children with lower eyelid trichiasis, suture method is simple and can be performed under local anesthesia in collaboration with children, but with a higher relapse rate, some patients required reoperation;partial resection of eyelashes can be more thoroughly solve the problem of pediatric eyelid trichiasis with low recurrence rate, but children need to be under general anesthesia with some of big risk. So partial resection of eyelashes is unsuitable for using in clinical practice widely and can be used in special cases.

Effects of Xue-Bi-Jing combined with forsythia suspension on the liver gene expression levels of rats with sepsis model / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study the effects of "XUE BI JING plus LIANQIAO" injection on gene expression levels of rats with sepsis model.</p><p><b>METHODS</b>One hundred and twenty rats were randomly divided into sham operation group, sepsis model group, Te-neng group and "XUE BI JING plus forsythia suspension" group. The sepsis model of rats was prepared by "CLP" method. Tai neng group was treated by peritoneal injection Imipenem/ Cilastatin (0.18 g/kg); "XUE BI JING plus LIANQIAO" group was treated by peritoneal injection Imipenem/ Cilastatin (0.18 g/kg) plus "xue-bi-jing" (10 ml/kg) and "liang ge san" (18 g/200 g) by intragastric administration 2 times a day; the sham operation group and model group were treated by peritoneal injection of normal saline (10 ml/kg). The survival rates at 48h and 72h were observed for all groups. The gene expression levels of livers in all groups were detected by BiostarR-40s chip. The NCBI database was used to inquest Gene function and class.</p><p><b>RESULTS</b>The survival rates at 48h and 72h in "XUE BI JING+ forsythia suspension" group were 83.3% and 76.7% which were significantly higher than those (30.0% and 16.7%) in sepsis model group and those (60.0% and 33.3%) in Te-neng group (P < 0.01). Model group/control group have 305 differential expression genes with 159 up-regulation genes and 146 down-regulation genes. Tai-neng group/model group have 386 differential expression genes with 206 up-regulation genes and 180 down-regulation genes. "XUE BI JING plus forsythia suspension" group/model group have 342 differential expression genes with 102 up-regulation genes and 240 downregulation genes. The genes with up-regulation in model group/ control group and with down-regulation in"XUE BI JING plus forsythia suspension" group/model group were 24. The genes with down-regulation in the model group/ sham operation group and with up-regulation in "XUE BI JING plus forsythia suspension"group/model group were 16.</p><p><b>CONCLUSION</b>"XUE BI JING plus forsythia suspension" can reduce the mortality of rats with sepsis, which could be due to the expression of relative regulation genes.</p>

Dynamic changes of alpha-AR, beta1-AR and beta2-AR expression during hepatic fibrogenesis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the dynamic changes of a-AR, b1-AR and b2-AR expression in hepatic fibrosis.</p><p><b>METHODS</b>Rat hepatic fibrosis model was established by bile duct ligation (BDL). HE and Masson staining were used to determine hepatic fibrosis levels. Immunohistochemistry was applied to detect alpha -smooth muscle actin (alpha -SMA), a marker of hepatic stellate cell (HSC) activation; Western blot and real-time RT-PCR were used to measure the dynamic changes of alpha -AR, beta(1)-AR, beta(2)-AR expression on protein and mRNA levels, respectively, during the development of hepatic fibrosis.</p><p><b>RESULTS</b>(1) HE and Masson trichrome staining showed that the liver fibrosis models were established successfully. (2) At 1, 2, 3, 4 wk after BDL, alpha -SMA positive area density of the model group (10.58% +/- 1.75%, 24.14% +/- 2.02%, 29.74% +/- 2.59%, 34.28% +/- 2.01%) was significantly higher than that of the sham operation group (4.12% +/- 1.51%), P less than 0.01. (3) The expression of alpha -AR, beta(1)-AR, beta(2)-AR protein and mRNA was increased with the development of the hepatic fibrosis (P less than 0.05). (4) alpha -SMA expression was positively associated with alpha -AR, beta(1)-AR, beta(2)-AR, r values were 0.564, 0.753 and 0.606, respectively.</p><p><b>CONCLUSION</b>The expression of alpha -SMA is increased dramatically during the fibrosis, and is positively associated with the expression of alpha -AR, beta(1)-AR and beta(2)-AR.</p>

Effect of Salvia miltiorrhiza monomer IH764-3 on MMP-13 and TIMP-1 by downregulating the expression of focal adhesion kinase in hepatic stellate cell stimulated by H2O2 / 中国应用生理学杂志

<p><b>AIM</b>To investigate the effect of IH764-3 on the expression of MMP-13 and TIMP-1 by H2O2-stimulated hepatic stellate cell and the alteration of FAK during this process.</p><p><b>METHODS</b>The expression of MMP-13 and FAK mRNA was examined by RT-PCR. TIMP-1 mRNA was analyzed by in-situ hybridization. FAK and TIMP-1 were evaluated at protein level through Western blotting method.</p><p><b>RESULTS</b>Being incubated for 2 h, compared with control group, MMP-13 mRNA was upregulated by IH764-3, but TIMP-1 transcription was reduced in a dose-dependent manner, accompanied with the decrease of FAK mRNA. The expression of TIMP-1 and FAK protein in HSC also decreased after being exposed by IH764-3 for 24 h.</p><p><b>CONCLUSION</b>IH764-3 can induce the expression of MMP-13 and inhibit the expression of TIMP-1. Down-regulating the expression of FAK mRNA may be one of its mechanisms.</p>

Effects of matrix metalloproteinase-3, osteopontin, and tissue inhibitor of metalloproteinase-1 in the formation of cataract / 中南大学学报(医学版)

OBJECTIVE@#To investigate the expression and significance of matrix metalloproteinase-3 (MMP-3), osteopontin (OPN) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in human lens epithelial cells (LECs) of cataract.@*METHODS@#The MMP-3, OPN, and TIMP-1 expressions in LECs of anterior subcapsular cataract (31 cases), nuclear cataract (28 cases) and normal lens (12 cases) were detected by immunohistochemistry, respectively.@*RESULTS@#The MMP-3 expression in anterior subcapsular cataract was significantly higher than that in nuclear cataract and normal lens (chi2=31.49, 34.479; P=0.000); but there was no statistic significance between nuclear cataract and normal lens (chi2=2.449, P=0.118). The OPN expression in anterior subcapsular cataract was also significantly higher than that in nuclear cataract and normal lens (chi2=29.450, 15.889; P=0.000). There was no significant difference in the TIMP-1 expression among the 3 groups (P>0.05). Positive correlation was found between MMP-3 and OPN in LECs of anterior subcapsular cataract (r=0.381, P=0.035). But no significant correlation was found among MMP-3, OPN, and TIMP-1 (r=0.121, -0.289; P=0.516, 0.114).@*CONCLUSION@#Increased expression of MMP-3 and OPN and the expression imbalance between MMP-3 and TIMP-1 may play a critical role in the pathogenesis of anterior subcapsular cataract.

Immobilization of cells by macro-porous NaCS-PDMDAAC capsules and cultivation in shaking flask and bubble bioreactor / 生物工程学报

The membrane of sodium cellulose sulphate ( NaCS)-poly dimethyldiallylammonium chloride (PDMDAAC) microcapsule is compact and has low molecular weight cut-off, which would delay the mass transfer and affect the cell growth immobilized in the capsule. Macroporous NaCS-PDMDAAC microcapsules were prepared using the degradation of the starch by amylase in the membrane of the capsules. The pore size and the permeability in the membrane were improved obviously. As model cells, the Candida krusei CK1 and E. coli EC1 immobilized in the capsules were cultured in the shake flask and bubble column respectively. It was shown that the cell density immobilized in the microcapsules cultured in the bubble column was higher than that cultured in the shaking flask. It implied that the limiting factor of the cell growth in the capsule lied in the diffusion of the oxygen. Since the rate of the oxygen transporting across the membrane was greatly enhanced due to the enlarged pore size, the maximum cell density in the macroporous capsules was 20%-110% over than that in the standard capsules in the bubble column. However, the extent of E. coli cell density increasing was higher than that of the yeast, which may be due to the difference of the oxygen requirement between the two microbes.

Can tumor uptake Tc-99m MDP ?

Abstract. To explore the mechanism of technetium-99m-methylene diphosphonate (MDP) uptake within tumor through analyze a distribution of Tc-99m MDP in mice bearing tumor cell lines. Methods: The uptake of Tc-99m MDP was analyzed in seven human tumor cell lines ( SPC-A1 adenocarcinoma of lung cancer, Bcap-37 Breast cancer, T-24 Bladder cancer, SKOV3 Ovary carcinoma, Hela-229 Cervical carcinoma, SCI-OS Osteosarcoma, SCI-375 Melanoma) and mouse Lewis lung cancer cell line. They were transplanted into athymic mice, SCID nude mice and C57BL/6 mice, respectively. Approximately 10(6) cells of each cell line were injected subcutaneously into a right chest of mouse. After 4&5 weeks, the Tc-99m MDP scintigraphy were determined 6 hours after tail vein injection of 74MBq in 0.05ml every mouse. Result: Biodistribution and tumor uptake MDP was different in the various cell types investigated. According to the RegionRatio program of Siemens Power Macintosh 9500 Computer System, region of interests (RIOs) placed on a small part of the tumor and horizontal copied to left background (T/B) and thoracic spine (T/N) of mice in Tc-99m MDP imaging. The average cpm/pixel ratios were calculated by standardized uptake measure (SUM) and determined the tumor-positive value (T/B) greater than or equal to 1.2. T/B of cell lines were sorted from higher to lower as follows: SCI-OS, Lewis, SKOV3, SCI-375, T-24, SPC-A1, Bcap-37, Hela-229. T/N: SCI-OS, SKOV3, T-24, SCI-375, Lewis, SPC-A1, Bcap-37, Hela-229. The biodistribution data of 99Tcm-MDP in SPC-A1 tumor-bearing BALB/c nude mice were given as ID/g and represent the meansñSD (n=13) in 30 hours after injection of Tc-99m MDP. ID/g of major tissue were sorted from higher to lower as follows: thoracic spine, lumbar, ribs, kidneys, the center of tumor, the ulcer of tumor, the surrounding of tumor, lymph node, blood, lungs, heart, liver. Conclusions: Most of tumor can uptake Tc-99m MDP including human adenocarcinoma. The uptake rate in the center tissue of tumor is higher than other part of tumor. The uptake rate of tumor is higher than non-skeletal tissue unless kidneys. It maybe connected with necrosis or calcification of tumor. Calcium and phosphorus ions were seen frequently in larger tumor. Not only it was caused by fibrous scar and/or surrounding tissues of granuloma but also intra-tumor coagulation and liquefaction necrosis